# Dietary menaquinone-9 supplementation does not influence bone tissue quality or bone mineral density during skeletal development in mice

## TL;DR

This study found that adding menaquinone-9 to the diet of young mice did not improve bone quality or density during development.

## Contribution

The study is the first to investigate the effects of dietary menaquinone-9 on bone health in mice.

## Key findings

- Dietary MK9 supplementation did not significantly affect bone tissue quality or bone mineral density in mice.
- Higher concentrations of MK4 in bone did not correlate with improved skeletal properties.
- Female mice showed reduced cross-sectional area and moment of inertia with higher MK9 doses, but not males.

## Abstract

Vitamin K has been implicated in skeletal health because vitamin K-dependent proteins are present in bone tissue. While there are multiple forms of vitamin K, most research has focused on phylloquinone, which is found mainly in plant-based foods, and its metabolite menaquinone-4 (MK4). However, there are additional forms of vitamin K that are bacterially produced that appear to influence bone health but have not yet been studied extensively. Herein, we evaluated the effects of menaquinone-9 (MK9), a bacterially produced form of vitamin K, on bone tissue quality and density in young mice. Four-week-old male (n = 32) and female (n = 32) C57BL/6 mice were supplemented with 0.06 mg/kg diet or 2.1 mg/kg diet of MK9 for 12 wk. During week 11, a subgroup of mice (n = 7/sex/group) received daily deuterium-labeled MK9 to trace its metabolic fate in bone. Liver MK4 and MK9 were significantly higher in mice fed 2.1 mg MK9/kg compared to those receiving 0.06 mg MK9/kg, regardless of sex (all p ≤ .017). MK4 was the only vitamin K form detected in bone, with 63%-67% of skeletal MK4 in mice fed 2.1 mg MK9/kg derived from deuterium-labeled MK9. Femoral tissue strength, maximum bending moment, section modulus, and BMD did not differ significantly between diet groups in either sex (all p ≥ .083). Cross-sectional area (p = .003) and moment of inertia (p = .001) were lower in female mice receiving 2.1 mg MK9/kg compared to those receiving 0.06 mg MK9/kg, but no differences were found in male mice. Higher bone MK4 concentrations did not correlate with higher bone tissue quality or density. Despite dietary MK9 being a dietary precursor to MK4 in bone, dietary MK9 supplementation did not affect bone tissue quality or BMD during skeletal development.